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使用敲除细胞株进行验证重组RabMAb

重组Anti-ROCK2抗体[EPR7141(B)] (ab125025)

Reviews (4)Q&A (4)References (47)
Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
  • Immunocytochemistry/ Immunofluorescence - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
  • Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
  • Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
  • Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
  • Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
  • OI-RD Scanning - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
  • Anti-ROCK2 antibody [EPR7141(B)] (ab125025)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR7141(B)] to ROCK2
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 555 Alexa Fluor® 647 Carrier Free

概述

  • 产品名称

    Anti-ROCK2抗体[EPR7141(B)]
    参阅全部 ROCK2 一抗

  • 描述

    兔单克隆抗体[EPR7141(B)] to ROCK2

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, ICC/IFmore details
    不适用于: IHC-P

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide within Human ROCK2 (C terminal). The exact sequence is proprietary.

  • 阳性对照

    • HepG2, A549, HeLa, HEK293, L6, RAW264.7 and A673 cell lysates; Human kidney tissue, Mouse and Rat brain tissue lysate.

  • 常规说明

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab125025于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB (3)
1/10000 - 1/50000. Detects a band of approximately 161 kDa (predicted molecular weight: 161 kDa).
ICC/IF
1/100 - 1/250.
说明
WB
1/10000 - 1/50000. Detects a band of approximately 161 kDa (predicted molecular weight: 161 kDa).
ICC/IF
1/100 - 1/250.
应用说明
Is unsuitable for IHC-P.

靶标

  • 功能

    Regulates the assembly of the actin cytoskeleton. Promotes formation of stress fibers and of focal adhesion complexes. Plays a role in smooth muscle contraction.

  • 序列相似性

    Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family.
    Contains 1 AGC-kinase C-terminal domain.
    Contains 1 PH domain.
    Contains 1 phorbol-ester/DAG-type zinc finger.
    Contains 1 protein kinase domain.
    Contains 1 REM (Hr1) repeat.

  • 翻译后修饰

    Phosphorylated upon DNA damage, probably by ATM or ATR.

  • 细胞定位

    Cytoplasm. Cell membrane. Cytoplasmic, and associated with actin microfilaments and the plasma membrane.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • coiled-coil-containing protein kinase 2 antibody
    • KIAA0619 antibody
    • p164 ROCK 2 antibody
    • p164 ROCK-2 antibody
    • Rho associated coiled coil containing protein kinase 2 antibody
    • Rho associated protein kinase 2 antibody
    • Rho associated, coiled coil containing protein kinase II antibody
    • Rho kinase 2 antibody
    • Rho-associated antibody
    • Rho-associated protein kinase 2 antibody
    • ROCK 2 antibody
    • Rock II antibody
    • Rock2 antibody
    • ROCK2_HUMAN antibody
    • Rock2m antibody
    • ROK alpha antibody
    • ROKalpha antibody
    see all

图片

  • Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
    Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: ROCK2 knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: A10 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab125025 observed at 165 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab125025 was shown to specifically react with ROCK2 when ROCK2 knockout samples were used. Wild-type and ROCK2 knockout samples were subjected to SDS-PAGE. ab125025 and ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
    Immunocytochemistry/ Immunofluorescence - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)

    Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling ROCK2 with purified ab125025 at 1/250. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab7291 anti-Tubulin (mouse mAb) primary and ab150120 (AlexaFluor®594 goat anti-mouse) secondary both at 1/1000 dilution. Nuclei were counterstained with DAPI (blue).

    For negative control 1, rabbit primary antibody and ab150120 (anti-mouse) secondary antibody were used. For negative control 2, ab7291 (mouse primary antibody) was used followed by ab150077 (anti-rabbit secondary antibody).

  • Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
    Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
    All lanes : Anti-ROCK2 antibody [EPR7141(B)] (ab125025) at 1/10000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ROCK2 knockout HeLa cell lysate
    Lane 3 : HepG2 cell lysate
    Lane 4 : A549 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 161 kDa
    Observed band size: 175 kDa why is the actual band size different from the predicted?



    Lanes 1- 4: Merged signal (red and green). Green - ab125025 observed at 175 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab125025 was shown to react with ROCK2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265679 (knockout cell lysate ab257643) was used. Wild-type HeLa and ROCK2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab125025 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
    Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
    All lanes : Anti-ROCK2 antibody [EPR7141(B)] (ab125025) at 1/50000 dilution

    Lane 1 : HeLa whole cell lysate
    Lane 2 : HEK293 whole cell lysate
    Lane 3 : Mouse brain tissue lysate
    Lane 4 : Rat brain tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 161 kDa
    Observed band size: 161 kDa



    Blocking and Diluting buffer 5% NFDM/TBST
  • Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
    Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
    All lanes : Anti-ROCK2 antibody [EPR7141(B)] (ab125025) at 1/10000 dilution (unpurified)

    Lane 1 : Hela cell lysate
    Lane 2 : L6 cell lysate
    Lane 3 : A673 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 161 kDa

  • Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
    Western blot - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
    Anti-ROCK2 antibody [EPR7141(B)] (ab125025) at 1/10000 dilution + RAW264.7 whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 161 kDa
    Observed band size: 161 kDa



    Blocking and Diluting buffer 5% NFDM/TBST
  • OI-RD Scanning - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
    OI-RD Scanning - Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
    Anti-ROCK2 antibody [EPR7141(B)] (ab125025)

文献 (47)

发表研究结果有使用 ab125025？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab125025 被引用在 47 文献中.

  • Liu H  et al. Rho A/ROCK1 signaling-mediated metabolic reprogramming of valvular interstitial cells toward Warburg effect accelerates aortic valve calcification via AMPK/RUNX2 axis. Cell Death Dis 14:108 (2023). PubMed: 36774349
  • Chang H  et al. Highly-expressed lncRNA FOXD2-AS1 in adipose mesenchymal stem cell derived exosomes affects HaCaT cells via regulating miR-185-5p/ROCK2 axis. Adipocyte 12:2173513 (2023). PubMed: 36775902
  • Huang S  et al. Corydalis decumbens Alleviates the Migration, Phagocytosis, and Inflammatory Response of Macrophages. Evid Based Complement Alternat Med 2023:7000477 (2023). PubMed: 36874618
  • Sambandam A  et al. Obligate role for Rock1 and Rock2 in adult stem cell viability and function. Heliyon 9:e14238 (2023). PubMed: 36950615
  • Marcadis AR  et al. Rapid cancer cell perineural invasion utilizes amoeboid migration. Proc Natl Acad Sci U S A 120:e2210735120 (2023). PubMed: 37075074
View all Publications for this product

客户评价及客户问答

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1-8 of 8 Abreviews or Q&A

Western blot abreview for Anti-ROCK2 antibody [EPR7141(B)]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Pig Cell lysate - whole cell (HEART)
Gel Running Conditions
Reduced Denaturing
Loading amount
5 µg
Specification
HEART
Blocking step
Licor as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 24°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Nov 04 2016

Western blot abreview for Anti-ROCK2 antibody [EPR7141(B)]

 Average
Abreviews
abreview image
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
Human Tissue lysate - whole (CNS tissue)
Specification
CNS tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Aug 14 2014

Western blot abreview for Anti-ROCK2 antibody [EPR7141(B)]

 Good
Abreviews
abreview image
Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
Mouse Tissue lysate - whole (brain tissue)
Specification
brain tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 May 19 2014

Immunohistochemistry (Frozen sections) abreview for Anti-ROCK2 antibody [EPR7141(B)]

 Average
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain (hippocampus))
Specification
Brain (hippocampus)
Fixative
Paraformaldehyde
Blocking step
10% goat serum, 2% horse serum, 1%BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Sofia Santos

Verified customer

提交于 Apr 25 2013

Question

Thank you for your reply! I hope your holiday was nice and relaxing!
We have already ordered the ab125025 antibody, in part because of your technical responses.
To ensure that we use the antibody properly, is it possible to provide me with the more detailed Western Blot protocol used by your scientists with this antibody? Specifically, the amount and type of blocking and antibody solution (milk or other?), buffer (PBS(T) or TBS(T)), primary antibody incubation time and temperature, secondary incubation time, etc? If you can provide this, that would be great!

Read More
Answer

Thank you for contacting us.

As we do not beliefe in "one-fits-all" protocols, and as we do not have any advise on this datasheet, which means that most standard protocols used in our customer labs should be fine. We only give out general advise on e.g. blocking agents when we (or our customers) came along significant problems like when the antibody will work only with 3% BSA or needs 1h incubation only. Please keep in mind that every antibody needs it individual optimization, too.

Nonetheless, for your convenience, I have attached our standard protocol to this email.



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Question

Inquiry: To whom it may concern, I am interested to know if your anti-ROCK2 rabbit monoclonal antibody (ab125025) was tested for cross-reactivity to ROCK1? On a similar subject, another ROCK2 antibody (ab136442) does not cross-react with ROCK1. Will you please tell me how this was determined? Kindly,

Read More
Answer

Thank you for contacting us and for your patience.

I contacted the laboratory to require more information about how the specificity of ab136442 was tested. I can confirm, that the immunogen sequence of this antibody shows an identity of 75.5% to ROCK1. The specificity was tested performing a dotBlot.

However, I can confirm that the immunogen sequence for ab125025 is not contained in the amino acid sequence of ROCK1 at all. This makes this product very specific for the detection of ROCK2.

For this reason and because the ab125025 is covered by our six month guarantee for more applications (WB, IHC-P, ICC/IF) and species (Mouse, Rat, Human) I would recommend using the ab125025.

If you want to perform IP experiments you might be eligible for the AbTrial with ab125025. You can read more about our AbTrial offer here:

https://www.abcam.com/index.html?pageconfig=resource&rid=11998

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Question

I think I wrote it in the wrong way,
I would like to test an antibody that can detect ROCK (PCP pathway) in westerns and immuno staining. I does not need to inhibit the signalling itself.
Which one can you recommend?

Read More
Answer

Thank you for contacting us.

All the antibodies are well characterized, please choose based on your requirements. e.g polyclonal/ monoclonal, Rabbit/mouse/sheep etc.

The catalogue numbers are:

ab13660, ab45171, ab56661, ab58305, ab125025

ab71598, ab66310, ab10549, ab97592, ab85612

When selecting check the Abreview and choose the antibody which have more Abreview and publications.

You may also interested in testing the antibody. Please click the below link for more info.

https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Question

I have been using some of antibodies a lot for Immuno-Staining and Western Blotting.
At the moment I am interested in looking at a functional ROCK antibody and its regulation during PCP / Wnt signalling.
Is there an antibody in your product catalogue you can recommend ?
Thanks for your help,
Constanze

Read More
Answer

Thank you for contacting us.

We unfortunately do not have anti ROCK antibodies available that can be used for functional assays e.g. activation or inhibition of ROCK. I would suggest searching the similar antibody clone in publications. We have 6-7 monoclonal antibodies in catalogue however we do not know whether these are being used in any functional assay. I am sorry we are unable to help with this particular ab.

I hope this information is nevertheless helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

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